Colony-stimulating factors (CSFs) are secreted glycoproteins which bind to receptor proteins on the surfaces of hemopoietic stem cells and thereby activate intracellular signaling pathways which can cause the cells to proliferate and differentiate into a specific kind of blood cell (usually white blood cells). In humans there are three CSF genes, CSF1 which encodes macrophage CSF (M-CSF), CSF2, which encodes the granulocytes macrophage CSF (GM-CSF), and CSF3, which encodes the granulocyte CSF (G-CSF).
Hamilton, Nature Reviews 8 (2008), pp 533-544 reports that depletion of CSFs have a therapeutic benefit in many inflammatory and/or autoimmune diseases and that there are numerous antibody therapies in clinical development targeted to CSFs for therapy of inflammation.
Neupogen®(Filgrastim) is a heterologously produced human G-CSF produced by Amgen for enhancing white blood cell concentration in cancer patients being treated with chemotherapy.
G-CSF has been indicated in chronic inflammatory autoimmune diseases, such as type II hypersensitivity responses, including rheumatoid arthritis.
MicroRNA-155 is induced during the macrophage inflammatory response (O'Connell et al., PNAS 104 (5) pp 1604-9).
WO2008/017126 refers antisense compounds which target the granulocyte colony-stimulating factor (G-CSF), and the use of such compounds for the treatment of pulmonary disease. G-CSF protein has been developed as a therapeutic agent for increasing white blood cell counts, and can enhance the immune system's ability to raise a Th-2 response that can decrease Th-1 mediated inflammatory responses, for example in Crohn's disease.
G-CSF
Granulocyte colony-stimulating factor (G-CSF) is a colony-stimulating factor hormone. It is a glycoprotein, growth factor or cytokine produced by a number of different tissues to stimulate the bone marrow to produce granulocytes and stem cells. G-CSF then stimulates the bone marrow to release them into the blood. It also stimulates the survival, proliferation, differentiation, and function of neutrophil precursors and mature neutrophils.
It is playing importance in inflammatory joint diseases as G-CSF-deficient mice are protected from acute and chronic arthritis. Reduced severity was associated with blunted mobilization of granulocytic cells from the bone marrow and less cellular infiltrate and cellular activation in inflamed joints. It has also been demonstrated that G-CSF blockade in established collagen-induced arthritis in WT mice markedly reduces disease manifestations and is as effective as tumor necrosis factor blockade. G-CSF plays a critical role in driving joint inflammation and G-CSF is a potential therapeutic target in inflammatory joint diseases, such as rheumatoid arthritis (Lawlor et al., PNAS, 2004).
Worsening of Psoriasis afte treatment with G-CSF have been reported (Feliu et al., JNCI, 1997) indicating a role for G-CSF in the pathogenesis of Psoriasis.
M-CSF
Macrophage colony-stimulating factor, or M-CSF, is a secreted cytokine which influences hemopoietic stem cells to differentiate into macrophages or other related cell types. Also the macrophage-colony stimulating factor, M-CSF supports osteoclast formation (Yoshida et al. Nature 345: 442-444, 1990). Osteoclasts mediate bone readsorption. Osteoclasts are multinucleated cells differentiating from haemopoietic cells (Walker, Science 190: 784-785, 1975) and they share a common stem cell with monocyte-macrophage lineage cells (Ash et al., Nature 283: 669-670, 1980). The differentiation of osteoclast precursors into mature multinucleated osteoclasts requires different factors including hormonal and local stimuli (Walker, Science 190: 784-785,1975) and living bone and bone cells have been shown to play a critical role in osteoclast development (Hagenaars et al., Bone Miner 6: 179-189,1989). Osteoblastic or bone marrow stromal cells are also required for osteoclast differentiation and one of the factors produced by these cells that supports osteoclast formation is macrophage-colony stimulating factor, M-CSF (Yoshida et al., Nature 345: 442-444, 1990).
Thus, there remains a need in the art to identify new agents and methods for preventing or treating osteolysis or cancer metastasis, including osteolytic bone metastases. Metabolic bone diseases associated with relatively increased osteoclast activity, includes endocrinopathies (including hypercortisolism, hypogonadism, primary or secondary hyperparathyroidism, hyperthyroidism), hypercalcemia, deficiency states (including rickets/osteomalacia, scurvy, malnutrition), chronic diseases (including malabsorption syndromes, chronic renal failure (including renal osteodystrophy), chronic liver disease (including hepatic osteodystrophy)), drugs (including glucocorticoids (glucocorticoid-induced osteoporosis), heparin, alcohol), and hereditary diseases (including osteogenesis imperfecta, homocystinuria), cancer, osteoporosis, osteopetrosis, inflammation of bone associated with arthritis and rheumatoid arthritis, periodontal disease, fibrous dysplasia, and/or Paget's disease.
M-CSF plays a more general role in formation of cancer metastasis. Studies of M-CSF null mutant mice demonstrated that M-CSF plays an important role in mammary tumor progression to metastasis. M-CSF regulates these processes through the recruitment and regulation of macrophages, cells that become associated with mammary tumors and the terminal end buds at the end of the growing ducts. This phenomenon suggests that the tumors subvert normal developmental processes to allow invasion into the surrounding stroma, a process that gives the tumor access to the vasculature and consequently the promotion of metastasis. In addition, soluble M-CSF secreted from the tumor acts to divert antitumor macrophage responses and suppresses the differentiation of mature tumor-antigen-presenting dendritic cell (Lin et al., J. Exp.Med, 2002).
Chemokine (C-C Motif) Ligand 2 (CCL2) (Alt.: Monocyte Chemoattractant Protein-1 (MCP-1))
The chemoattractant cytokines, termed as chemokines, are a large family of low molecular weight proteins that share the ability to stimulate directed cell migration [Schall, Cytokine 3:165-183 (1991); Murphy, Rev Immun 12:593-633 (1994)]. Chemokines have been implicated as important mediators of allergic, inflammatory and autoimmune disorders and diseases, such as asthma, atherosclerosis, glomerulonephritis, pancreatitis, restenosis, rheumatoid arthritis, diabetic nephropathy, pulmonary fibrosis, multiple sclerosis, and transplant rejection. Accordingly, the use of antagonists of chemokine function may help reverse or halt the progression of these disorders and diseases.
With few exceptions, chemokines have four conserved cysteine residues that form disulfide bonds within the chemokine proteins. Two major chemokine subfamilies have been classified based on the chromosomal localization of the chemokine genes and the relative position of the first two cysteine residues (Van Collie et al., Cytokine Growth Factor Rev 10:61-86 (1999)). Monocyte chemoattractant protein-1 (CCL-2) is a member of the C-C class of the beta chemokine family and one of the key factors involved in the initiation of inflammation. CCL-2 is typically secreted in the prevalent forms, 9 and 13 kDa, respectively, as a result of differential O-glycosylation. It triggers chemotaxis and transendothelial migration of monocytes to inflammatory lesions by interacting with the membrane CC chemokine receptor 2 (CCR2) in monocytes (O'Hayre et al., 2008). CCL-2 is secreted by fibroblasts, endothelial cells, vascular smooth muscle cells, monocytes, T cells, and other cell types that mediate the influx of cells to sites of inflammation (Conti and DiGioacchino, 2001). CCL-2 expression has been observed in a large number of tissues during inflammation-dependent disease progression, including atherosclerosis (Shin et al., 2002), arthritis (Taylor et al., 2000) and cancer (O'Hayre et al., 2008). In these cases, the influx of macrophages into these tissues has been suggested to exacerbate the diseases. Thus, the expression of CCL-2, which is likely to be critical for fighting infectious disease, must be tightly regulated.
CCL-2 In Diseases
Elevated expression of CCL-2 has been observed in a number of chronic inflammatory diseases [Proost et al., Int J Clin Lab Res 26:211-223 (1996); Taub, D. D. Cytokine Growth Factor Rev 7:355-376 (1996)] including, but not limited to, rheumatoid arthritis [Robinson et al., Clin Exp Immunol 101:398-407 (1995); Hosaka et al., ibid. 97:451-457 (1994); Koch et al., J Clin Invest 90:772-779 (1992); Villiger et al., J Immunol 149:722-727 (1992)], asthma [Hsieh et al., J Allergy Clin Immunol 98:580-587 (1996); Alam et al., Am J Respir Crit Care Med 153:1398-1404 (1996); Kurashima et al., J Leukocyte Biol 59:313-316 (1996); Sugiyama et al., Eur Respir J 8:1084-1090 (1995)], and atherosclerosis [Yla-Herttuala et al., Proc Natl Acad Sci USA 88:5252-5256 (1991); Nelken et al., J Clin Invest 88:1121-1127 (1991)].
CCL-2 appears to play a significant role during the early stages of allergic responses because of its ability to induce mast cell activation and LTC4 release into the airway, which directly induces AHR (airways hyper-responsiveness) [Campbell et al., J Immunol 163:2160-2167 (1999)].
CCL-2 has been found in the lungs of patients with idiopathic pulmonary fibrosis and is thought to be responsible for the influx of mononuclear phagocytes and the production of growth factors that stimulate mesenchymal cells and subsequent fibrosis [Antoniades et al., Proc Natl Acad Sci USA 89:5371-5375 (1992)]. In addition, CCL-2 is also involved in the accumulation of monocytes in pleural effusions implicated in both Mycobacterium tuberculosis infection and malignancy [Strieter et al., J Lab Clin Med 123:183-197 (1994)].
CCL-2 has also been shown to be constitutively expressed by synovial fibroblasts from rheumatoid arthritis patients, and its levels are higher in rheumatoid arthritis joints compared to normal joints or those from other arthritic diseases [Koch et al., J Clin Invest 90:772-779 (1992)]. These elevated levels of CCL-2 are probably responsible for the monocyte infiltration into the synovial tissue. CCL-2 also plays a critical role in the initiation and development of atherosclerotic lesions. CCL-2 is responsible for the recruitment of monocytes into atherosclerotic areas, as shown by immunohistochemistry of macrophage-rich arterial wall [Yla-Herttuala et al., Proc Natl Acad Sci USA 88:5252-5256 (1991); Nelken et al., J Clin Invest 88:1121-1127 (1991)] and anti-CCL-2 antibody detection [Takeya et al., Human Pathol 24:534-539 (1993)]. LDL-receptor/CCL-2-deficient and apoB-transgenic/CCL-2-deficient mice show significantly less lipid deposition and macrophage accumulation throughout their aortas compared with wild-type CCL-2 strains [Alcami et al., J Immunol 160:624-633 (1998); Gosling et al., J Clin Invest 103:773-778 (1999); Gu et al., Mol. Cell. 2:275-281 (1998); Boring et al., Nature 394:894-897 (1998). Other inflammatory diseases marked by specific site elevations of CCL-2 include multiple sclerosis (MS), glomerulonephritis, and stroke. Together, these findings infer CCL-2 as a therapeutic target in the treatment of inflammatory disease and strongly suggest that the discovery and development of novel compounds that block or down-regulate CCL-2 activity would be highly beneficial in treating inflammatory diseases.
Interleukine-6 (IL-6)
IL-6 is a multifunctional cytokine originally identified as a T cell-derived factor that causes the terminal maturation of antigen-stimulated immature B-cells into immunoglobulin-producing plasma cells [Hirano T, Taga T, Nakano N, Yasukawa K, Kashiwamura S, Shimizu K, et al. Proc Natl Acad Sci USA 1985; 82: 5490-4]. A number of cell types produce IL-6, including T-cells, B-cells, monocytes, fibroblasts, keratinocytes, endothelial cells, mesangial cells and bone marrow stroma cells [Kawano M, Hirano T, Matsuda T, Taga T, Horii Y, Iwato K, et al. Nature 1988; 332: 83-5]. IL-6 also has a wide range of responder cells, including B-cells, T-cells, hepatocytes, hemotopoietic precursor cells, neural cells, epidermal keratinocytes, mesangial cells, and osteoclasts [Adachi et al. Current Pharmaceutical Design, 2008, 14, 1217-1224]. IL-6 functions as an immune regulator, acute phase protein inducer, cell differentiation factor, cell growth factor, and bone metabolism regulator against these effector cells. Additionally, IL-6 induces C-reactive protein (CRP) and serum amyloid A (SAA) on hepatocytes [Adachi et al. Current Pharmaceutical Design, 2008, 14, 1217-1224]. Both of these proteins are important markers of inflammation and are used clinically in monitoring patients suffering from inflammatory conditions. Recently, IL-6 has been implicated in the balance of Th17 and regulatory T cells has made the novel focus in immunology [Tato C M, O'Shea J J. Nature 2006; 441: 166-8.]. Given that aberrant helper T cell regulation is observed in chronic inflammatory states in humans, this action may confer superiority to anti-IL-6 treatments over approaches targeting other inflammatory cytokines.
IL-6 In Inflammatory Diseases
IL-6 is one of the key regulators of the inflammatory responses and induces the final maturation of B-cells into immunoglobulin-producing cells [Adachi et al. Current Pharmaceutical Design, 2008, 14, 1217-1224]. Owing to these properties, IL-6 is a pivotal molecule in the pathogenesis of several chronic inflammatory diseases, such as Castleman's disease, rheumatoid arthritis (RA), juvenile idiopathic arthritis, and Crohn's disease [Adachi et al. Current Pharmaceutical Design, 2008, 14, 1217-1224]. These diseases are often refractory to conventional therapies such as corticosteroids and immunosuppressants. Additionally, IL-6 overproduction plays an important pathological role in several neoplasms, including high-grade multiple myelomas [17-19] and malignant mesotheliomas [20, 21]. The paraneoplastic syndrome of mesothelioma including immunosuppression, cachexia, thrombocytosis, and amyloidosis, is related to IL-6 overproduction [Nakano et al. Br J Cancer 1998; 77: 907-912; Fitzpatrick et al. Am J Respir Cell Mol Biol 1995; 12: 455-60]. As such, anti-IL-6 treatment may both alleviate the clinically devastating paraneoplastic syndrome and suppress tumor growth. Thus, therapeutics targeting IL-6 show high potential for the treatment of inflammatory conditions and malignancies.
There is therefore a need to develop agents which can either down-regulate expression of genes such as M-CSF, G-CSF, CCL-2 and IL-6, or up-regulate genes such as M-CSF, GCSF, CCL-2 and IL-6 for use in the treatment of diseases wherein the modulation of expression of those factors will be beneficial. As is apparent herein, it may also be desirable for such agents to modulate the expression of other immune-related genes, such as Bcl2l1, Cd40, Nos2, Socs1, Stat1, and Cxcr3.